| Objective:As the prevalence of diabetes increases year by year,the susceptibility to ischemiareperfusion of diabetic patients has attracted extensive attention.Whether the strategy that pretreatment with inhaled anesthetics to reduce ischemia-reperfusion injury during surgery,which is commonly used in clinic,is still applicable to diabetic patients deserves further study.The purpose of this study was to investigate the effect and intrinsic mechanism of inhaled anesthetic sevoflurane on diabetic renal ischemia-reperfusion injury,and the effect of Foxo1 inhibitor AS1842856 on the protective effect of sevoflurane to diabetic renal ischemia-reperfusion.Methods:1.C57 Wild mice were divided into 3 groups with 10 rats in each group,which were given sham operation,renal ischemia-reperfusion and sevoflurane-preconditioning renal ischemia-reperfusion treatment respectively.Then to observe the result of blood glucose,blood creatinine,urea nitrogen and Pas staining of pathological sections.2.C57 mice were continuously injected with streptozotocin for 5 days to establish type 1 diabetes model.Then these mice were divided into three groups with 10 rats in each group,and were given sham operation,renal ischemia-reperfusion and sevoflurane-preconditioning renal ischemia-reperfusion respectively.To observe the changes of blood glucose,blood creatinine,urea nitrogen levels and the Pas staining injury results of pathological sections in different groups of diabetic mice.3.In the group of sevoflurane-precongditioning diabetic renal ischemia-reperfusion,before the sevoflurane pretreatment,Group AS1842568 and sevoflurane-preconditioning renal ischemia-reperfusion in diabetic rat(D+I+AS group)were treated with Foxo1 inhibitor AS1842856,and the other groups were given the same amount of methylcellulose.To observe the expression levels of blood glucose,blood creatinine,urea nitrogen and Pas staining of pathological sections in mice pretreated with AS1842856.Western Blot was used to detect the protein expression of p-Akt,p-Foxo1,Foxo1 and Caspase-3 in each diabetic mice group,and RT-qPCR was used to detect the mRNA expression of Akt,Foxo1,Caspase-3 and the downstream apoptotic genes Bim,Fas,glycosides gene G6 Pase,PEPCK,and insulin transcription factors MafA,NeuroD1 and Pdx of Foxo1.Results:1.In wild mice,sevoflurane pretreatment can reduce blood creatinine(P<0.01),urea nitrogen levels(P<0.05)and PAS staining injury score(P<0.05),and exert the protective effect of renal ischemia-reperfusion.2.After comorbiding with diabetes,there was no change in blood creatinine and urea nitrogen levels in the sevoflurane-preconditioning renal ischemia reperfusion group,and the PAS injury score did not decrease too(P>0.05).The protective effect of renal ischemiareperfusion disappeared.3.In diabetic mice,the protein expression of p-Akt,p-Foxo1 and Caspase-3 in the sevoflurane-preconditioning ischemia-reperfusion group was unchanged.Meanwhile,the mRNA expression of Akt,Foxo1,Caspase-3 and the downstream genes of Foxo1: Bim,Fas,G6 Pase,PEPCK,MafA,NeuroD,and Pdx1 also showed no difference(P>0.05).However,Giving AS1842568 in advance before the pretreatment with sevoflurane,the result showed that the blood creatinine(P<0.01)and urea nitrogen levels(P<0.05)decreased,and the Pas staining injury score decreased(P<0.01);the protein expression of p-Akt and p-Foxo1 increased(P<0.01),Caspase-3 protein expression decreased(P <0.01),Foxo1 protein and mRNA expression were not different(P>0.05);the mRNA expression of Akt and Pdx1 increased(P<0.01),Caspase-3 and the downstream gene of Foxo1:Bim,Fas,G6 Pase,PEPCK,MafA and NeuroD were down-regulated(P<0.01).Conclusion:The inhaled anesthetic sevoflurane has the protective effect of renal ischemiareperfusion,which can protect the kidney from the ischemia-reperfusion injury during surgery in C57 mice.However,when with diabetes,its protective effect disappeared,which may be related to the inhibition of PI3 K / Akt / Foxo1 signaling pathway.Administration of Foxo1 inhibitor AS1842568 combined with dephosphorylated Foxo1,can down-regulated the expression of Foxo1 and it’s downstream regulatory factors,which will reshape the protective effect of sevoflurane on renal ischemia-reperfusion. |
